1168. Are the Staphylococcus aureus Isolates that Recolonize Infants in a NICU Following Successful Decolonization the Same or Different from the Initial Colonizing Isolate?

Abstract Background Staphylococcus aureus is an important pathogen of infants in a neonatal intensive care unit (NICU). Colonization precedes infection and decolonization may prevent infection. The origin of colonizing organisms may be the NICU environment or personnel or visitors. We have observed infants who became recolonized after successful decolonization. The purpose of this study was to determine the proportion of infants who become recolonized with the same strain or a different strain. Methods Eligible infants were consecutive infants who 1. were colonized with methicillin-susceptible S. aureus and were successfully decolonized with topical mupirocin ointment (nares and umbilicus) as evidenced by 2 or more consecutive negative weekly surveillance cultures (in the absence of a course of systemic antibiotics with activity against MSSA), 2. subsequently became recolonized, and 3. the pair of isolates was available for analysis. Isolates were analyzed by staphylococcal protein A (spa) typing and pairs with concordant spa types were subjected to whole genome sequencing (WGS; Illumina MiSeq) and phylogenetic analyses. Pairs of isolates with fewer than 25 single nucleotide polymorphism differences were considered closely related. Results There were 19 occurrences of MSSA recolonization in 17 infants following 2-6 (median, 2) negative weekly intervening surveillance cultures. Based upon spa typing (that identified 19 spa types), in 11 (58%) there was a concordant spa type and in 8 (42%) there was a discordant spa type. Of the 11 pairs of isolates with concordant spa types that were compared after WGS, 10 were closely related resulting overall in recolonization with a closely related strain in 53% of episodes. Conclusion Among MSSA colonized infants who become recolonized after successful decolonization, the recolonizing strain is the same as the original strain in over half of cases. In such cases the source is more likely to be a visitor than the NICU environment or staff. The possibility that some cases classified as recolonization were in fact persistent low level colonization or carriage in another body site not detected by surveillance cultures cannot be excluded. Disclosures Anne-Catrin Uhlemann, MD, PhD, Merck (Grant/Research Support)

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The molecular epidemiology of Staphylococcus aureus of bovine mastitis origin

Veterinarski arhiv ◽

10.24099/vet.arhiv.0808 ◽

2021 ◽

Vol 91 (1) ◽

pp. 1-10

Author(s):

Tawheed Ahmad Shafi ◽

◽

Baljinder K. Bansal ◽

Shukriti Sharma ◽

Dhiraj K. Gupta ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Molecular Epidemiology ◽

Protein A ◽

Bovine Mastitis ◽

Subclinical Mastitis ◽

Staphylococcal Protein A ◽

Spa Typing ◽

New Novel ◽

Spa Types ◽

High Diversity

The present study was aimed to understand the molecular epidemiology of Staphylococcus aureus (54 isolates), isolated from 422 milk samples obtained from 108 subclinical mastitis affected cows (CMT positive ≥1+ in at least one quarter). The molecular epidemiology of Staphylococcus aureus was studied using coagulase (coa) gene polymorphism, 16S-23S ribosomal spascer (RS-PCR) polymorphism and Staphylococcal protein A (Spa) typing. Staphylococcus aureus produced 7 coagulase genotypes and 8 RS genotypes respectively. Coagulase genotype GTIII (730 bp) was the most prevalent (35 strains) followed by GTV (900 bp, 7 strains) and GTIV (800 bp, 4 strains), whereas RS genotypes GTA accounted for the highest number of strains (31 strains), followed by GTB (11strains), GTH (4 strains) and GTE (3 strains). Coagulase genotype CTIII (730 bp) showed the highest diversity, as isolates within it produced 5 RS genotypes, the majority of them belonging to the RS genotype GTA (29 out of 31 strains). Forty out of 54 Staphylococcus aureus samples isolated in this study were correctly typed by spa typing, and were assigned to 21 known spa types, and one new novel spa type t18462. The study revealed high diversity within Staphylococcus aureus strains, consisting of 7 coagulase genotypes, 8 RS genotypes and 22 spa types.

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1164. Acquisition of Methicillin-Susceptible Staphylococcus aureus in the Neonatal Intensive Care Unit: Molecular Comparison of Relatedness of Isolates

Open Forum Infectious Diseases ◽

10.1093/ofid/ofab466.1357 ◽

2021 ◽

Vol 8 (Supplement_1) ◽

pp. S673-S674

Author(s):

Dalia F Eid ◽

Angela Gomez-Simmonds ◽

Vitaliya Boyar ◽

Anne-Catrin Uhlemann ◽

Lorry G Rubin

Keyword(s):

Neonatal Intensive Care ◽

Protein A ◽

Illumina Miseq ◽

Healthcare Personnel ◽

Control Group ◽

Staphylococcal Protein A ◽

Spa Typing ◽

Exact Test ◽

Molecular Comparison ◽

Spa Type

Abstract Background In the NICU MSSA infections occur frequently and cause morbidity and mortality. Colonization is a risk factor for infection. Optimal infection prevention strategies await a more complete understanding of acquisition and transmission. To investigate possible transmission, we studied whether newly MSSA colonized infants share a strain with another contemporaneously colonized infant. Methods This is a prospective observational study in a level IV NICU from April through November 2019. Infants had weekly MSSA nasal surveillance cultures. Isolates from newly MSSA colonized infants and other infants colonized with MSSA during the same/previous week were subjected to staphylococcal protein A (spa) typing; most pairs with a concordant (CC) spa type were analyzed by whole genome sequencing (WGS; Illumina MiSeq). Pairs of isolates with a CC spa type and < 25 single nucleotide polymorphism differences on WGS were considered closely related (CC pairs). A control group consisted of pairs of isolates from a newly colonized infant with one randomly chosen colonized infant with a discordant (DC) spa type during the same/previous week. The medical records were reviewed for staff member (SM) and room assignment. Fischer’s exact test was used to compare proportions. Results Isolates from 60/68 consecutive newly MSSA colonized infants and 111/133 comparison infants were available for spa typing. Of these 60 infants, 23 (38 %) had a CC spa type with another infant colonized during the same/previous week. Of 18 isolate pairs from infants with a CC spa type that were subjected to WGS, 12 (67%) pairs of isolates were closely related. 7/12 (58 %) of CC pairs had a SM in common compared to 2/13 (15 %) in the DC pair groups, p=0.04. 2/12 (17 %) of CC pairs shared a room compared to 2/13 (15 %) pairs in the DC group, p=1.0. Conclusion Among newly MSSA colonized infants at least 25% are colonized with an isolate closely related to that of another colonized infant indicating likely infant to infant transmission. WGS is more discriminatory than spa typing for MSSA. Given the lack of commonality of room assignment and the commonality of SM assignment, a possible role of healthcare personnel in MSSA transmission should be further investigated. Disclosures Anne-Catrin Uhlemann, MD, PhD, Merck (Grant/Research Support)

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Multilocus Sequence Typing and Staphylococcal Protein A Typing Revealed Novel and Diverse Clones of Methicillin-Resistant Staphylococcus aureus in Seafood and the Aquatic Environment

Journal of Food Protection ◽

10.4315/0362-028x.jfp-16-260 ◽

2017 ◽

Vol 80 (3) ◽

pp. 476-481 ◽

Cited By ~ 6

Author(s):

V. Murugadas ◽

C. Joseph Toms ◽

Sara A. Reethu ◽

K. V. Lalitha

Keyword(s):

Staphylococcus Aureus ◽

Aquatic Environment ◽

Multilocus Sequence Typing ◽

Methicillin Resistant Staphylococcus Aureus ◽

Protein A ◽

Staphylococcal Protein A ◽

Spa Typing ◽

Methicillin Resistant ◽

Staphylococcal Protein ◽

Mrsa Strains

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) has been a global health concern since the 1960s, and isolation of this pathogen from food-producing animals has been increasing. However, little information is available on the prevalence of MRSA and its clonal characteristics in seafood and the aquatic environment. In this study, 267 seafood and aquatic environment samples were collected from three districts of Kerala, India. Staphylococcal protein A (spa) typing and multilocus sequence typing (MLST) was performed for 65 MRSA strains isolated from 20 seafood and aquatic environment samples. The MRSA clonal profiles were t657-ST772, t002-ST5, t334-ST5, t311-ST5, t121-ST8, t186-ST88, t127-ST1, and two non-spa assignable strains. Whole spa gene sequence analysis along with MLST confirmed one strain as t711-ST6 and another as a novel MRSA clone identified for the first time in seafood and the aquatic environment with a t15669 spa type and a new MLST profile of ST420-256-236-66-82-411-477. The MRSA strains were clustered into five clonal complexes based on the goeBURST algorithm, indicating high diversity among MRSA strains in seafood and the aquatic environment. The novel clone formed a separate clonal complex with matches to three loci. This study recommends large-scale spa typing and MLST of MRSA isolates from seafood and the aquatic environment to determine the prevalence of new MRSA clones. This monitoring process can be useful for tracing local spread of MRSA isolates into the seafood production chain in a defined geographical area.

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Molecular characterization of clonal lineage and staphylococcal toxin genes fromS. aureusin Southern Nigeria

PeerJ ◽

10.7717/peerj.5204 ◽

2018 ◽

Vol 6 ◽

pp. e5204 ◽

Cited By ~ 3

Author(s):

Funmilola A. Ayeni ◽

Werner Ruppitsch ◽

Franz Allerberger

Keyword(s):

Protein A ◽

High Potential ◽

Clinical Samples ◽

Staphylococcal Protein A ◽

Clonal Lineage ◽

Clinical Strains ◽

Enterotoxin Genes ◽

Southern Nigeria ◽

Spa Types ◽

Spa Type

BackgroundStaphylococcus aureusis a human colonizer with high potential for virulence, and the spread of the virulent strains from the colonized hosts to non-carriers in the community is on the increase. However, there are few reports on comprehensive analysis of staphylococcal enterotoxin (SE) genes with clonal lineage inS. aureusin Africa. This is essential because of diversity of cultures and habits of the people. This study analyzed spa types and enterotoxin genes inS. aureusstrains previously isolated from the human nostrils, poultry and clinical samples in Southern Nigeria.MethodsForty-sevenS. aureusisolates were obtained from humans nostrils (n = 13), clinical strains (n = 21) and poultry (n = 13) from previous studies in Southern Nigeria. The strains were analyzed formecAgene, selected toxins genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq, ser, seu)and Panton-Valentine leukocidin (PVL) gene(lukS-PV/lukF-PV)by PCR. Population structures of the strains were detected by Staphylococcal protein A (spa) typing.ResultsTwenty different spa types were obtained with the highest percentages, 17% observed inspatype t091 from clinical, nasal and poultry samples while t069 was the most prevalent spa type in poultry. Two MRSA were only detected in human strains. The poultry strains had the highest occurrence of SE genes (18%) followed by nasal strains (15%) and clinical strains (10%). Eighty-nine percent of all tested isolates harbored at least one SE gene;seowas the most prevalent (34%) followed byseg(30%) andsea(21%), whilesec, seeandsejwere absent in all strains. Spa type t355 was associated withlukS-PV/lukF-PVgene and complete absence of all studied SE.Sea, seq, seb, sekwere associated with spa type t069;seawas associated with t127 whilesepwas associated with spa type t091. There were coexistences ofseo/segandsei/seg.ConclusionsThe higher carriage of staphylococci enterotoxin genes by the nasal and poultryS. aureusstrains suggests a high potential of spread of staphylococcal food poisoning through poultry and healthy carriers in the community. This is the first report of high occurrence of staphylococcal enterotoxins genes in poultry from Nigeria.

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Molecular typing of Staphylococcus aureus collected from a Major Hospital in Amman, Jordan

The Journal of Infection in Developing Countries ◽

10.3855/jidc.3676 ◽

2014 ◽

Vol 8 (04) ◽

pp. 441-447 ◽

Cited By ~ 6

Author(s):

Dana A Bazzoun ◽

Houda H Harastani ◽

Asem A Shehabi ◽

Sima T Tokajian

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Agents ◽

Protein A ◽

University Hospital ◽

Epidemiological Typing ◽

Spa Typing ◽

Study Results ◽

Typing Methods ◽

Healthcare Associated ◽

Spa Types

Introduction: Over the past decade methicillin-resistant Staphylococcus aureus (MRSA) has been recognized as a major cause of healthcare associated infections. Recently, however, epidemiology of this pathogen has changed drastically with the emergence of new clones in the community. Efficient epidemiological typing methods are essential to monitor and limit the occurrence and spread of epidemic clones. Methodology: A total of sixty S. aureus isolates were collected from the Jordan University hospital in Amman-Jordan. All isolates were characterized using Staphylococcus protein A (spa) typing and pulsed-field gel electrophoresis (PFGE). Samples were tested for their susceptibility patterns against seven antimicrobial agents and for their potential to form biofilms. Results: spa typing showed that spa type t044 was the most common representing 28% of the isolates studied and 38% of the MRSA population. PFGE revealed fourty-six pulsotypes among the sixty tested isolates clustering similar spa types together. The predominant resistance was detected against levofloxacin, chloramphenicol and clindamycin. One MSSA isolate typed as spa t955 showed biofilm formation potential through protein deposition.. Conclusion: The study results are based on one hospital, but the findings of this and other studies conducted in the region indicate that there is an urgent need for standardized surveillances combined with the application of well-validated typing methods to assess the occurrence of MRSA and to control its spread.

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Molecular characterization of clonal lineage and Staphylococcal toxin genes from S. aureus in Southern Nigeria

10.7287/peerj.preprints.26535v1 ◽

2018 ◽

Author(s):

Funmilola Ayeni ◽

Werner Ruppitsch ◽

Franz Allerberger

Keyword(s):

Protein A ◽

Food Poisoning ◽

Staphylococcal Protein A ◽

Toxin Genes ◽

Serious Infections ◽

Southern Nigeria ◽

Population Structures ◽

Toxic Shock Syndrome Toxin ◽

Spa Types ◽

Spa Type

Background. The pathogenic role of S. aureus as causative agent of serious infections and food poisoning is on the increase. However, there are few reports on comprehensive analyses of toxins and staphylococcal enterotoxin (SE) genes in S. aureus in Africa. This study analyzed spa types and toxin genes in S. aureus obtained from our previous studies in Southern Nigeria. Methods. Forty-seven non-duplicate S. aureus isolates were obtained from humans (n = 34) and poultry (n = 13) from previous studies in Southern Nigeria. The strains were analyzed for mecA, selected toxins genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq, ser, seu), TSST and lukS-PV/lukF-PV by PCR. Population structures of the strains were detected by Staphylococcal protein A (spa) typing. Results. Twenty three percent of all isolates (47) carried the Panton-Valentine leukocidin (PVl) gene. Two MRSA were detected. Twenty different spa types were obtained, with the highest percentages, 17% belonging to spa type t091 was observed in 4 states from clinical, nasal and poultry samples while t069 is the most prevalent type in poultry. Eighty-nine percent of the all tested isolates harbored at least one staphylococcus enterotoxin. Seo was the most prevalent SE (34%) followed by seg (30%) and sea (21%), while toxic shock syndrome toxin (TSST), seb,sec,,see, sej, sel, sem, and ser, seu were absent in all strains. Spa type t355 was associated with the PVl and complete absence of all studied SE. Sea, seq, seb, sek were associated with spa type 069; t127 was associated with sea while sep was associated with spa type t091. There was coexistence of seo/seg and sei/seg. Conclusions. We detected a high incidence of enterotoxins and PVl encoding genes in these potential staphylococcal reservoir. Specific toxin genes were observed in particular spa types.

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Novel spa and Multi-Locus Sequence Types (MLST) of Staphylococcus Aureus Samples Isolated from Clinical Specimens in Korean

Antibiotics ◽

10.3390/antibiotics8040202 ◽

2019 ◽

Vol 8 (4) ◽

pp. 202 ◽

Cited By ~ 1

Author(s):

Yae Sung Mun ◽

You Jin Hwang

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Susceptibility ◽

Bacterial Infections ◽

Protein A ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Sequence Types ◽

Spa Types ◽

Spa Type

Staphylococcus aureus bacteremia is one of the most frequent and severe bacterial infections worldwide. The increased incidence of S. aureus infections with a diverse pattern of S. aureus protein A (spa) types across different geographic regions is a global challenge. This study investigated a novel spa type of methicillin-resistant S. aureus in a clinically isolated specimen. A total of 109 clinical S. aureus samples were subjected to 19 sets of antimicrobial susceptibility testing using the Kirby–Bauer disk diffusion method. Molecular typing was performed with S. aureus protein A (spa) and multi-locus sequence types (MLST) via polymerase chain reaction and sequencing. The methicillin-resistant S. aureus samples in our study accounted for 55.05% (60/109) of the total. A novel spa type was detected in five (5/60) strains. This gh22 isolate was identified in antimicrobial susceptibility tests of 15 kinds of antibiotics. Antibiotic resistance genes included mecA, TEM, aac(6′)-aph(2”), ermA, and tetM. Eleven S. aureus samples were classified as t2460, t338, t324, t693, five unknown spa types (new spa types), and undefined MLST (novel MLST). We report a high prevalence rate of t2460 methicillin-resistant S. aureus samples in our country. Additionally, novel spa gh22, MLST ST4613, and clonal compact CC5-type strains (T1:M1:B1:B1:M1:E1:K1, r26:r17:r34:r34:r17:r13:r16, mlst;1:4:1:4:559:495:10) showing multidrug resistance were identified among S. aureus samples.

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Coagulase Typing and spa Typing of Methicillin Resistant Staphylococcus aureus: Relatedness Among Patients' and Carrier Strains

Bangladesh Journal of Medical Microbiology ◽

10.3329/bjmm.v2i1.21780 ◽

2016 ◽

Vol 2 (1) ◽

pp. 3-7

Author(s):

Sadia Afroz ◽

ABM Bayezid Hossain ◽

Afzalunnessa Binte Lutfor ◽

Naima Muazzam

Keyword(s):

Staphylococcus Aureus ◽

Wound Infection ◽

Methicillin Resistant Staphylococcus Aureus ◽

Hospital Staff ◽

Spa Typing ◽

Methicillin Resistant ◽

Mrsa Strains ◽

Spa Types ◽

Spa Type

This study was carried out to determine the types of Methicillin-Resistant Staphylococcus aureus (MRSA) by two typing systems, Coagulase typing and spa typing and to identify relationship between types of MRSA strains isolated from patients and hospital staff carriers. A total of 40 MRSA strains, 33 from cases of wound infection and 7 from anterior nares of hospital staffs were investigated. Coagulase typing showed that all the MRSA strains isolated from patients were coagulase type VI and all the strains isolated from carriers were coagulase type VII. In spa typing, seven spa types were detected such as S4, S6, S7, S8, S9, S10 and S11. Predominant spa type in patients was S7 detected in 15 (45.45%) MRSA isolates, followed by S4 in 8 (24.24%) strains and S6 in 5(15.15%) strains. Other spa types detected in patients in small numbers were S8 (3.03%), S9 (3.03%), S10 (6.06%) and S11 (3.03%). While in strains of MRSA isolated from carriers, only two types S4 and S9 were detected. Both the spa types identified from carriers were also detected in a total of 9 (27.2%) of MRSA strains isolated from patients.Bangladesh J Med Microbiol 2008; 02 (01): 3-7DOI: http://dx.doi.org/10.3329/bjmm.v2i1.21780

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Molecular characterization of clonal lineage and Staphylococcal toxin genes from S. aureus in Southern Nigeria

10.7287/peerj.preprints.26535 ◽

2018 ◽

Author(s):

Funmilola Ayeni ◽

Werner Ruppitsch ◽

Franz Allerberger

Keyword(s):

Protein A ◽

Food Poisoning ◽

Staphylococcal Protein A ◽

Toxin Genes ◽

Serious Infections ◽

Southern Nigeria ◽

Population Structures ◽

Toxic Shock Syndrome Toxin ◽

Spa Types ◽

Spa Type

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1217. Staphylococcus Protein A (spa) Typing Demonstrates Genetic Heterogeneity of Methicillin-Susceptible Staphyloccus aureus (MSSA) in a Neonatal Intensive Care Unit (NICU)

Open Forum Infectious Diseases ◽

10.1093/ofid/ofy210.1050 ◽

2018 ◽

Vol 5 (suppl_1) ◽

pp. S369-S369

Author(s):

Alexandra Hill-Ricciuti ◽

Samantha Ferguson ◽

Wenjing Geng ◽

Stephania Stump ◽

Maria Messina ◽

...

Keyword(s):

Genetic Heterogeneity ◽

Neonatal Intensive Care ◽

Local Community ◽

Protein A ◽

Infection Prevention And Control ◽

Multiple Sources ◽

Spa Typing ◽

Incident Cases ◽

Spa Types ◽

Spa Type

Abstract Background In the NICU, MSSA is a more prevalent pathogen than MRSA, but optimal infection prevention and control strategies for MSSA are not yet well understood. There are likely multiple routes of MSSA acquisition given its role as normal flora and its detection in the anovaginal tract of pregnant women. We describe the molecular epidemiology of MSSA in our NICU during a yearlong surveillance effort. Methods Included infants were hospitalized in a university-affiliated level III-IV NICU from January to December 2017 (1032 admissions) and had positive clinical and/or surveillance cultures for MSSA. Infants admitted at ≥7 days of age were screened for MSSA colonization by culturing the anterior nares and three skin sites. All infants in the NICU were screened twice monthly. Spa typing was performed to genetically characterize isolates. Results During the study period, MSSA was identified in 187 infants (18 at admission, 145 by twice monthly surveillance, and 24 from clinical cultures). In all, 269 MSSA isolates (245 surveillance and 24 clinical isolates) from 166 infants were spa typed. Sixty-two MSSA spa types were identified; 31 (50%) were each detected in only one infant. The incidence of the nine most common spa types is shown (Figure 1); t279 (13%), t002 (8%), and t1451 (6%) had the highest incidence. t1451 and t571 belong to ST398, a common MSSA clone in the local community. The epidemiology of spa types varied; e.g., incident cases of t279 was detected in 10 months, t1451 was detected in 6 months and t148 in 3 months. Among the 14 sets of twins and triplets with MSSA isolates, 12 (86%) shared the same spa type as their sibling(s). Of the 58 infants with >1 MSSA isolate, 12 (21%) acquired new spa-types. No spa type(s) predominated in the 19 episodes of invasive infections. In 6 infants with both colonizing and invasive isolates, colonizing and invasive isolates were the same spa type(s) in 5. Conclusion Spa typing demonstrated that MSSA isolates in our NICU exhibited substantial genetic heterogeneity. While these data do not elucidate acquisition route(s), they suggest infants are acquiring MSSA from multiple sources, likely including family members and the local community. Ongoing sequencing studies are examining common spa types to further understand transmission dynamics. Disclosures A. C. Uhlemann, Merck: Investigator, Grant recipient

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